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Android fat is a surrogate measure of visceral obesity in the truncal region. Both visceral adiposity and oxidative stress (OS) are linked to cardiometabolic risk factors and clinical cardiovascular disease. However, whether body fat distribution (android vs gynoid) is associated with OS remains unknown. We hypothesized that increased android fat will be associated with greater OS. Body fat distribution and markers of OS, including plasma levels of reduced (cysteine and glutathione) and oxidized (cystine and glutathione disulfide) aminothiols, were estimated in 711 volunteers (67% female, 23% black, mean age 48 ± 11) enrolled in the Emory Georgia Tech Predictive Health study. At 1 year, 498 subjects had repeat testing. At baseline, anthropometric and fat distribution indexes, including body mass index, waist circumference, weight/hip ratio, and android and gynoid fat mass correlated with lower plasma concentrations of glutathione and higher cystine levels indicative of higher OS. At 1 year, the change in android but not gynoid fat mass or body mass index negatively correlated with the change in the plasma glutathione level after adjustment for cardiovascular risk factors. Increased body fat, specifically android fat mass, is an independent determinant of systemic OS, and its change is associated with a simultaneous change in OS, measured as plasma glutathione. In conclusion, our findings suggest that excess android or visceral fat contributes to the development of cardiovascular disease through modulating OS.
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Distribuição da Gordura Corporal , Doenças Cardiovasculares/metabolismo , Estresse Oxidativo , Absorciometria de Fóton , Adulto , Índice de Massa Corporal , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/prevenção & controle , Feminino , Humanos , Estilo de Vida , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: Redox status and inflammation are important in the pathophysiology of numerous chronic diseases. Epidemiological studies have linked vitamin D status to a number of chronic diseases. We aimed to examine the relationships between serum 25-hydroxyvitamin D [25(OH)D] and circulating thiol/disulphide redox status and biomarkers of inflammation. DESIGN: This was a cross-sectional study of N = 693 adults (449 females, 244 males) in an apparently healthy, working cohort in Atlanta, GA. Plasma glutathione (GSH), cysteine (Cys) and their associated disulphides were determined with high-performance liquid chromatography, and their redox potentials (Eh GSSG and Eh CySS) were calculated using the Nernst equation. Serum inflammatory markers included interleukin-6 (IL-6), interleukin-8 (IL-8) and tumour necrosis factor-α, assayed on a multiplex platform, and C-reactive protein (CRP), assayed commercially. Relationships were assessed with multiple linear regression analyses. RESULTS: Serum 25(OH)D was positively associated with plasma GSH (ß ± SE: 0·002 ± 0·0004) and negatively associated with plasma Eh GSSG (ß ± SE: -0·06 ± 0·01) and Cys (ß ± SE: -0·01 ± 0·003) (P < 0·001 for all); statistical significance remained after adjusting for age, gender, race, percentage body fat and traditional cardiovascular risk factors (P = 0·01-0·02). The inverse relationship between serum 25(OH)D and CRP was confounded by percentage body fat, and full adjustment for covariates attenuated serum 25(OH)D relationships with other inflammatory markers to nonstatistical significance. CONCLUSIONS: Serum 25(OH)D concentrations were independently associated with major plasma thiol/disulphide redox systems, suggesting that vitamin D status may be involved in redox-mediated pathophysiology.
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Cisteína/sangue , Glutationa/sangue , Vitamina D/sangue , Feminino , Dissulfeto de Glutationa/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Oxirredução , Vitamina D/análogos & derivadosRESUMO
BACKGROUND: The authors investigated the relationship between self-reported sleep characteristics and brachial artery flow-mediated dilation (FMD) in a community-based population. Previous studies document that sleep apnea may be related to endothelial dysfunction but disagree whether subjective reports of sleep may also reflect such associations. METHODS: In 684 subjects (32% male) aged between 37 and 60 years enrolled in the Emory-Georgia Tech Predictive Health Institute study, the authors measured reported sleep characteristics using the Epworth Sleepiness Scale and the Pittsburgh Sleep Quality Index (PSQI) along with cardiovascular risk factors. Endothelial function was assessed using brachial artery FMD. Multivariate analysis of covariance was used to adjust for various cardiovascular risk factors including age, race, gender, smoking, hypertension, diabetes and body mass index. RESULTS: Lower brachial artery FMD values were correlated with higher Epworth Sleepiness Scale scores (P = 0.0275), even after adjustment for risk factors (P = 0.03). Total PSQI score was unrelated to brachial artery FMD. However, lower sleep quality (PSQI component 1) was associated with lower brachial artery FMD (multivariate P = 0.038), and participants who coughed or snored during sleep also had lower brachial artery FMD (6.24% ± 3.42%) compared with those who did not (6.92% ± 4.30%) (P = 0.056). This difference remained significant after adjustment for risk factors (P = 0.03). CONCLUSIONS: In a community-based population, our analysis indicates a significant association between sleepiness and snoring assessed by questionnaires and endothelial function. Simple subjective reports about individuals' sleep may be highly revealing indicators of endothelial function impairment and thus important indicators of cardiovascular disease risk.
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Endotélio Vascular/fisiologia , Autorrelato , Sono/fisiologia , Adulto , Velocidade do Fluxo Sanguíneo/fisiologia , Artéria Braquial/fisiologia , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Vigilância da População/métodos , Fatores de Risco , Autorrelato/normas , Inquéritos e Questionários/normas , Vasodilatação/fisiologiaRESUMO
INTRODUCTION: The acute respiratory distress syndrome (ARDS), affects up to 150,000 patients per year in the United States. We and other groups have demonstrated that bone marrow derived mesenchymal stromal stem cells prevent ARDS induced by systemic and local administration of endotoxin (lipopolysaccharide (LPS)) in mice. METHODS: A study was undertaken to determine the effects of the diverse populations of bone marrow derived cells on the pathophysiology of ARDS, using a unique ex-vivo swine preparation, in which only the ventilated lung and the liver are perfused with autologous blood. Six experimental groups were designated as: 1) endotoxin alone, 2) endotoxin + total fresh whole bone marrow nuclear cells (BMC), 3) endotoxin + non-hematopoietic bone marrow cells (CD45 neg), 4) endotoxin + hematopoietic bone marrow cells (CD45 positive), 5) endotoxin + buffy coat and 6) endotoxin + in vitro expanded swine CD45 negative adherent allogeneic bone marrow cells (cultured CD45neg). We measured at different levels the biological consequences of the infusion of the different subsets of cells. The measured parameters were: pulmonary vascular resistance (PVR), gas exchange (PO2), lung edema (lung wet/dry weight), gene expression and serum concentrations of the pro-inflammatory cytokines IL-1ß, TNF-α and IL-6. RESULTS: Infusion of freshly purified autologous total BMCs, as well as non-hematopoietic CD45(-) bone marrow cells significantly reduced endotoxin-induced pulmonary hypertension and hypoxemia and reduced the lung edema. Also, in the groups that received BMCs and cultured CD45neg we observed a decrease in the levels of IL-1ß and TNF-α in plasma. Infusion of hematopoietic CD45(+) bone marrow cells or peripheral blood buffy coat cells did not protect against LPS-induced lung injury. CONCLUSIONS: We conclude that infusion of freshly isolated autologous whole bone marrow cells and the subset of non-hematopoietic cells can suppress the acute humoral and physiologic responses induced by endotoxemia by modulating the inflammatory response, mechanisms that do not involve engraftment or trans-differentiation of the cells. These observations may have important implications for the design of future cell therapies for ARDS.
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Células da Medula Óssea/citologia , Transplante de Medula Óssea , Lesão Pulmonar/terapia , Doença Aguda , Animais , Células da Medula Óssea/metabolismo , Citocinas/sangue , Modelos Animais de Doenças , Endotoxinas/toxicidade , Regulação da Expressão Gênica , Antígenos Comuns de Leucócito/genética , Antígenos Comuns de Leucócito/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/etiologia , Lesão Pulmonar/patologia , Suínos , Transplante AutólogoRESUMO
AIMS: Arterial stiffening may lead to hypertension, greater left ventricular after-load and adverse clinical outcomes. The underlying mechanisms influencing arterial elasticity may involve oxidative injury to the vessel wall. We sought to examine the relationship between novel markers of oxidative stress and arterial elastic properties in healthy humans. METHODS AND RESULTS: We studied 169 subjects (mean age 42.6 ± 14 years, 51.6% male) free of traditional cardiovascular risk factors. Indices of arterial stiffness and wave reflections measured included carotid-femoral Pulse Wave Velocity (PWV), Augmentation Index (Aix) and Pulse Pressure Amplification (PPA). Non-free radical oxidative stress was assessed as plasma oxidized and reduced amino-thiol levels (cysteine/cystine, glutathione/GSSG) and their ratios (redox potentials), and free radical oxidative stress as derivatives of reactive oxygen metabolites (dROMs). Inflammation was assessed as hsCRP and interleukin-6 levels. The non-free radical marker of oxidative stress, cystine was significantly correlated with all arterial indices; PWV (r=0.38, p<0.001), Aix (r=0.35, p<0.001) and PPA (r=-0.30, p<0.001). Its redox potential, was also associated with PWV (r=0.22, p=0.01), while the free radical marker of oxidative stress dROMS was associated with Aix (r=0.25, p<0.01). After multivariate adjustment for age, gender, arterial pressure, height, weight, heart rate and CRP, of these oxidative stress markers, only cystine remained independently associated with PWV (p=0.03), Aix (p=0.01) and PPA (p=0.05). CONCLUSIONS: In healthy subjects without confounding risk factors or significant systemic inflammation, a high cystine level, reflecting extracellular oxidant burden, is associated with increased arterial stiffness and wave reflections. This has implications for understanding the role of oxidant burden in pre-clinical vascular dysfunction.
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Artérias/patologia , Rigidez Vascular , Adulto , Envelhecimento , Proteína C-Reativa/metabolismo , Doenças Cardiovasculares/patologia , Cistina/química , Elasticidade , Feminino , Humanos , Inflamação , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Oxirredução , Estresse Oxidativo , Fatores de RiscoRESUMO
Bone marrow-derived mesenchymal stem cells (BMDMSC) are emerging as a therapeutic modality in various inflammatory disease states, including acute lung injury (ALI). A hallmark of inflammation, and a consistent observation in patients with ALI, is a perturbation in the systemic redox environment. However, little is known about the effects of BMDMSC on the systemic redox status. The objective of the present study was to determine whether exogenously infused BMDMSC protect against endotoxin-induced oxidation of plasma cysteine (Cys) and glutathione (GSH) redox states. To determine the effect on the redox state if BMDMSC, mice received endotoxin intraperitoneally (1 mg/kg), followed by intravenous infusion of either 5 × 10(5) BMDMSC or an equal volume of saline solution. Control mice received intraperitoneal endotoxin followed by 5 × 10(5) lung fibroblasts given intravenously. Cys, cystine (CySS), GSH, and glutathione disulfide (GSSG) concentrations were determined by HPLC. Results showed sequential preservation of plasma Cys and GSH levels in response to BMDMSC infusion. The data show that BMDMSC infusion leads to a more reducing Cys and GSH redox state. The findings are the first to demonstrate that BMDMSC have antioxidant effects in vivo, and add to our understanding of the systemic effects of BMDMSC in lung injury.
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OBJECTIVE: Age-related changes in the larynx lead to significant voice impairment and reduced quality of life. There is a need for aged animal models that have practical generation times to study the fundamental changes and new therapeutics for the aging voice. The senescence accelerated prone mouse strain (SAMP) animals experience rapid aging without any experimental manipulation. The main objective of this study was to demonstrate the use of senescence accelerated mice to study aging in the larynx. STUDY DESIGN: Murine model. SETTING: Department of Animal Resources, Emory University. SUBJECTS AND METHODS: Larynges from five senescence accelerated prone mice, five normal aging senescence resistant mice, and five C57BL/6 mice were harvested and processed for paraffin sections. Histomorphometry was performed for assessment of collagen and hyaluronic acid distribution. In addition, frozen laryngeal tissue was harvested for transcriptional and translational assessment of collagen-1, using real-time polymerase chain reaction with specific primers and Western blots. Myofibroblast assessment was performed by immunostaining for the presence of alpha-smooth muscle actin. RESULTS: The deposition of collagen increased at six months of age in the SAMP vocal fold, and the level of collagen-1 mRNA increased with age. The myofibroblast protein alpha-smooth muscle actin was also found at a higher concentration in the SAMP vocal tissue. In contrast, the levels of hyaluronic acid in the vocal folds of SAMP mice decreased with age when compared to age-matched C57BL/6 mice. CONCLUSION: SAMP mice show accelerated, age-related changes in the vocal fold that were evident at as early as six months of age. The use of senescence accelerated mice offers promise as a model to study age-related laryngeal changes.
Assuntos
Envelhecimento/fisiologia , Laringe/fisiologia , Modelos Animais , Animais , Colágeno/metabolismo , Ácido Hialurônico/metabolismo , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , RNA Mensageiro/análiseRESUMO
The incidence of pulmonary fibrosis increases with age. Studies from our group have implicated circulating progenitor cells, termed fibrocytes, in lung fibrosis. In this study, we investigate whether the preceding determinants of inflammation and fibrosis were augmented with aging. We compared responses to intratracheal bleomycin in senescence-accelerated prone mice (SAMP), with responses in age-matched control senescence-accelerated resistant mice (SAMR). SAMP mice demonstrated an exaggerated inflammatory response as evidenced by lung histology. Bleomycin-induced fibrosis was significantly higher in SAMP mice compared with SAMR controls. Consistent with fibrotic changes in the lung, SAMP mice expressed higher levels of transforming growth factor-beta1 in the lung. Furthermore, SAMP mice showed higher numbers of fibrocytes and higher levels of stromal cell-derived factor-1 in the peripheral blood. This study provides the novel observation that apart from increases in inflammatory and fibrotic factors in response to injury, the increased mobilization of fibrocytes may be involved in age-related susceptibility to lung fibrosis.
Assuntos
Envelhecimento , Células da Medula Óssea/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Fibrose Pulmonar/metabolismo , Envelhecimento/efeitos dos fármacos , Animais , Antibióticos Antineoplásicos , Biomarcadores/metabolismo , Bleomicina , Condrócitos/metabolismo , Colágeno Tipo I/metabolismo , Colágeno Tipo IV/metabolismo , Modelos Animais de Doenças , Feminino , Células-Tronco Hematopoéticas/patologia , Hidroxiprolina/metabolismo , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Fibrose Pulmonar/fisiopatologia , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Several lines of evidence indicate that perturbations in the extracellular thiol/disulfide redox environment correlate with the progression and severity of acute lung injury (ALI). Cysteine (Cys) and its disulfide Cystine (CySS) constitute the most abundant, low-molecular-weight thiol/disulfide redox couple in the plasma, and Cys homeostasis is adversely affected during the inflammatory response to infection and injury. While much emphasis has been placed on glutathione (GSH) and glutathione disulfide (GSSG), little is known about the regulation of the Cys/CySS couple in ALI. The purpose of the present study was to determine whether endotoxin administration causes a decrease in Cys and/or an oxidation of the plasma Cys/CySS redox state (E(h) Cys/CySS), and to determine whether these changes were associated with changes in plasma E(h) GSH/GSSG. Mice received endotoxin intraperitoneally, and GSH and Cys redox states were measured at time points known to correlate with the progression of endotoxin-induced lung injury. E(h) in mV was calculated using Cys, CySS, GSH, and GSSG values by high-performance liquid chromatography and the Nernst equation. We observed distinct effects of endotoxin on the GSH and Cys redox systems during the acute phase; plasma E(h) Cys/CySS was selectively oxidized early in response to endotoxin, while E(h) GSH/GSSG remained unchanged. Unexpectedly, subsequent oxidation of E(h) GSH/GSSG and E(h) Cys/CySS occurred as a consequence of endotoxin-induced anorexia. Taken together, the results indicate that enhanced oxidation of Cys, altered transport of Cys and CySS, and decreased food intake each contribute to the oxidation of plasma Cys/CySS redox state in endotoxemia.
Assuntos
Lesão Pulmonar Aguda/sangue , Lesão Pulmonar Aguda/induzido quimicamente , Cisteína/sangue , Cistina/sangue , Endotoxinas/farmacologia , Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/patologia , Animais , Adesão Celular , Linhagem Celular , Citocinas/imunologia , Ingestão de Alimentos , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Endotoxinas/imunologia , Feminino , Glutationa/metabolismo , Dissulfeto de Glutationa/metabolismo , Humanos , Leucócitos/citologia , Leucócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Oxirredução , Mucosa Respiratória/citologia , Redução de PesoRESUMO
Several lines of evidence indicate that depletion of glutathione (GSH), a critical thiol antioxidant, is associated with the pathogenesis of idiopathic pulmonary fibrosis (IPF). However, GSH synthesis depends on the amino acid cysteine (Cys), and relatively little is known about the regulation of Cys in fibrosis. Cys and its disulfide, cystine (CySS), constitute the most abundant low-molecular weight thiol/disulfide redox couple in the plasma, and the Cys/CySS redox state (E(h) Cys/CySS) is oxidized in association with age and smoking, known risk factors for IPF. Furthermore, oxidized E(h) Cys/CySS in the culture media of lung fibroblasts stimulates proliferation and expression of transitional matrix components. The present study was undertaken to determine whether bleomycin-induced lung fibrosis is associated with a decrease in Cys and/or an oxidation of the Cys/CySS redox state and to determine whether these changes were associated with changes in E(h) GSH/glutathione disulfide (GSSG). We observed distinct effects on plasma GSH and Cys redox systems during the progression of bleomycin-induced lung injury. Plasma E(h) GSH/GSSG was selectively oxidized during the proinflammatory phase, whereas oxidation of E(h) Cys/CySS occurred at the fibrotic phase. In the epithelial lining fluid, oxidation of E(h) Cys/CySS was due to decreased food intake. Thus the data show that decreased precursor availability and enhanced oxidation of Cys each contribute to the oxidation of extracellular Cys/CySS redox state in bleomycin-induced lung fibrosis.
Assuntos
Cisteína/metabolismo , Cistina/metabolismo , Estresse Oxidativo/fisiologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Animais , Antibióticos Antineoplásicos/toxicidade , Bleomicina/toxicidade , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Ingestão de Alimentos , Espaço Extracelular/metabolismo , Feminino , Glutationa/metabolismo , Dissulfeto de Glutationa/metabolismo , Interleucina-6/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Oxirredução , Fibrose Pulmonar/patologiaRESUMO
BACKGROUND: Long-term success in lung transplantation is limited by obliterative bronchiolitis (OB), yet the mechanism for this disease is not well understood. Chemokine SDF-1 and its receptor, CXCR4, have been reported to be involved in several fibrogenic processes by recruiting inflammatory and fibroblast progenitor cells into injured tissues. We hypothesized that the SDF-1/CXCR4 axis also plays a role in the pathogenesis of OB. METHODS: Using the mouse heterotopic allogeneic airway transplant model, we transplanted mouse tracheas from BALB/c donors into C57BL/6 recipients. At Day 10 after transplant, we found high expression of SDF-1 in cells in the sub-epithelial layers of the allograft. Approximately 26% of cells infiltrating the allograft were CD45(+)CXCR4(+), as determined by flow cytometry analysis. RESULTS: Treatment of the recipients with a CXCR4 antagonist, TN14003, decreased cell infiltration into the grafts at Day 10 post-implantation. At Day 42, a significant reduction in luminal occlusion was found in the TN14003-treated animals compared with controls (57.40% vs 98.21%, p < 0.01). To demonstrate the relevance of the SDF-1/CXCR4 axis in OB, sections of lung tissue obtained from lung transplant patients with OB were examined for SDF-1 and CXCR4 expression. We found a higher number of CXCR4- and SDF-1-positive cells in samples from patients with OB as compared with normal lungs. CONCLUSIONS: These findings provide new insights into the mechanisms of lung chronic rejection and may lead to new intervention tools for the treatment of OB.
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Bronquiolite Obliterante/prevenção & controle , Quimiocina CXCL12/genética , Peptídeos/uso terapêutico , Complicações Pós-Operatórias/prevenção & controle , Receptores CXCR4/antagonistas & inibidores , Receptores CXCR4/genética , Traqueia/transplante , Animais , Bronquiolite Obliterante/genética , Quimiocina CXCL12/fisiologia , Citometria de Fluxo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Transplante Heterotópico , Transplante Homólogo/patologia , Transplante IsogênicoRESUMO
BACKGROUND: Results of treatment of severe T-lymphocyte immunodeficiencies by means of hematopoietic stem cell (HSC) transplantation have improved. T cell-depleted haploidentical transplantations are successful if there is no HLA-identical donor. Methods to remove T lymphocytes include addition of anti-CD52 antibodies and CD34(+) HSC selection. OBJECTIVE: Assessment of long-term immune function is important after these treatments. We looked at immune reconstitution in 36 survivors for more than 2 years after HSC transplantation for severe T-lymphocyte immunodeficiencies and compared engraftment quality between the 2 T-lymphocyte depletion methods. METHODS: Chimerism, T- and B-lymphocyte subsets, immunoglobulin levels, and specific antibody production at last follow-up were examined. The chi(2) (Fisher exact test) and Wilcoxon rank sum analyses were used to compare the groups. RESULTS: Nineteen patients received anti-CD52-treated and 19 anti-CD34-treated HSCs. More anti-CD52-treated patients had full donor myeloid chimerism (P = .025). All patients had full donor T-lymphocyte chimerism. There was no difference in donor B-lymphocyte chimerism, but significantly more anti-CD52-treated patients had class-switched memory B lymphocytes (P = .024), normal IgG levels, and normal responses to tetanus and Haemophilus influenzae type B vaccination. More anti-CD52-treated patients with common gamma chain or Janus-associated kinase 3 severe combined immunodeficiency had donor B lymphocytes. CONCLUSION: Long-term T-lymphocyte function is good with either treatment method, with a low incidence of graft-versus-host disease. The results imply more incomplete donor chimerism in anti-CD34-treated patients with less B-lymphocyte function.
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Anticorpos/uso terapêutico , Antígenos CD34/imunologia , Antígenos CD/imunologia , Antígenos de Neoplasias/imunologia , Glicoproteínas/imunologia , Transplante de Células-Tronco Hematopoéticas , Sistema Imunitário/fisiopatologia , Síndromes de Imunodeficiência/terapia , Linfócitos T , Linfócitos B , Antígeno CD52 , Infecções por Haemophilus/prevenção & controle , Haemophilus influenzae tipo b , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Sistema Imunitário/efeitos dos fármacos , Sistema Imunitário/patologia , Imunoglobulina G/sangue , Síndromes de Imunodeficiência/fisiopatologia , Memória Imunológica , Lactente , Subunidade gama Comum de Receptores de Interleucina/deficiência , Janus Quinase 3/deficiência , Estudos Longitudinais , Estudos Retrospectivos , Imunodeficiência Combinada Severa/imunologia , Imunodeficiência Combinada Severa/metabolismo , Imunodeficiência Combinada Severa/patologia , Índice de Gravidade de Doença , Toxoide Tetânico/uso terapêutico , Quimeras de Transplante , VacinaçãoRESUMO
Bone marrow-derived mesenchymal stem cells (BMDMSCs) appear to be important in repair of the chronic lung injury caused by bleomycin in mice. To determine effects of these BMDMSCs on an acute inflammatory response, we injected C57BL/6 mice intraperitoneally with 1 mg/kg endotoxin followed either by intravenous infusion of 5 x 10(5) BMDMSCs, the same number of lung fibroblasts, or an equal volume of normal saline solution. Lungs harvested 6, 24, and 48 h and 14 days after endotoxin showed that BMDMSC administration prevented endotoxin-induced lung inflammation, injury, and edema. Although we were able to detect donor cells in the lungs at 1 day after endotoxin, by 14 days no donor cells were detected. BMDMSC administration suppressed the endotoxin-induced increase in circulating proinflammatory cytokines without decreasing circulating levels of anti-inflammatory mediators. Ex vivo cocultures of BMDMSC and lung cells from endotoxemic animals demonstrated a bilateral conversation in which lung cells stimulated proliferation and migration of stem cells and suppressed proinflammatory cytokine production by lung cells. We conclude that BMDMSCs decrease both the systemic and local inflammatory responses induced by endotoxin. These effects do not require either lung engraftment or differentiation of the stem cells and are due at least in part to the production of stem cell chemoattractants by the lungs and to humoral and physical interactions between stem cells and lung cells. We speculate that mobilization of this population of BMDMSCs may be a general mechanism for modulating an acute inflammatory response.
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Células da Medula Óssea/metabolismo , Células-Tronco Mesenquimais/metabolismo , Síndrome de Resposta Inflamatória Sistêmica/prevenção & controle , Animais , Células da Medula Óssea/citologia , Comunicação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Citocinas/biossíntese , Endotoxinas , Feminino , Infusões Intravenosas , Lipopolissacarídeos/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pneumopatias/induzido quimicamente , Pneumopatias/patologia , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Edema Pulmonar/patologia , Síndrome de Resposta Inflamatória Sistêmica/induzido quimicamente , Receptor 4 Toll-Like/metabolismoRESUMO
Stromal cell-derived factor-1 (SDF-1) participates in mobilizing bone marrow-derived stem cells, via its receptor CXCR4. We studied the role of the SDF-1/CXCR4 axis in a rodent model of bleomycin-induced lung injury in C57BL/6 wild-type and matrix metalloproteinase (MMP)-9 knockout mice. After intratracheal instillation of bleomycin, SDF-1 levels in serum and bronchial alveolar lavage fluid increased. These changes were accompanied by increased numbers of CXCR4(+) cells in the lung and a decrease in a population of CXCR4(+) cells in the bone marrow that did not occur in MMP-9(-)/(-) mice. Both SDF-1 and lung lysates from bleomycin-treated mice induced migration of bone marrow-derived stem cells in vitro that was blocked by a CXCR4 antagonist, TN14003. Treatment of mice with TN14003 with bleomycin-induced lung injury significantly attenuated lung fibrosis. Lung tissue from patients with idiopathic pulmonary fibrosis had higher numbers of cells expressing both SDF-1 and CXCR4 than did normal lungs. Our data suggest that the SDF-1/CXCR4 axis is important in the complex sequence of events triggered by bleomycin exposure that eventuates in lung repair. SDF-1 participates in mobilizing bone marrow-derived stem cells, via its receptor CXCR4.
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Quimiocina CXCL12/metabolismo , Lesão Pulmonar , Receptores CXCR4/metabolismo , Animais , Bleomicina/toxicidade , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Movimento Celular/fisiologia , Feminino , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/patologia , Humanos , Técnicas In Vitro , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Metaloproteinase 9 da Matriz/deficiência , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Peptídeos/farmacologia , Fibrose Pulmonar/etiologia , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Receptores CXCR4/antagonistas & inibidoresRESUMO
RATIONALE: Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic lung disorder of unknown cause. Several studies suggest an association between Epstein-Barr virus pulmonary infection and the development of IPF. OBJECTIVES: To determine whether reduction of gamma-herpesvirus reactivation from latency would alter progressive lung fibrogenesis in an animal model of virus-induced pulmonary fibrosis. METHODS: IFN-gamma receptor-deficient (IFN-gammaR(-/-)) mice infected intranasally with murine gamma-herpesvirus 68 (MHV68) develop lung fibrosis that progresses for up to at least 180 days after initial infection. Viral replication during the chronic phase of infection was controlled by two methods: the administration of cidofovir, an antiviral drug effective at clearing lytic but not latent virus, and by using a mutant gamma-herpesvirus defective in virus reactivation from latency. MEASUREMENTS AND MAIN RESULTS: Ten percent of the asymptomatic MHV68-infected animals that received antiviral treatment beginning on Day 45 postinfection had severe pulmonary fibrosis compared with 40% of the control saline-treated animals. Absence of severe fibrosis was also observed in IFN-gammaR(-/-) mice infected with the defective reactivation mutant MHV68 v-cyclin stop. Decreased fibrosis was associated with lower levels of transforming growth factor-beta, vascular endothelial growth factor, and markers of macrophage alternative activation. When antiviral treatment was administered on Day 60 in symptomatic animals, survival improved from 20 to 80% compared with untreated symptomatic animals, but lung fibrosis persisted in 60% of the mice. CONCLUSIONS: MHV68-induced fibrosis is a result of viral lytic replication during chronic lung herpesvirus infection in mice. We speculate that antiviral therapy might help to control lung fibrosis in humans with IPF and associated herpesvirus infection.
Assuntos
Infecções por Herpesviridae/complicações , Fibrose Pulmonar/virologia , Receptores de Interferon/deficiência , Rhadinovirus/fisiologia , Infecções Tumorais por Vírus/complicações , Ativação Viral , Animais , Antígenos Virais/imunologia , Arginase/metabolismo , Biomarcadores/metabolismo , Líquido da Lavagem Broncoalveolar , Citocinas/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Técnica Indireta de Fluorescência para Anticorpo , Regulação Viral da Expressão Gênica , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Hidroxiprolina/metabolismo , Imunoensaio , Peptídeos e Proteínas de Sinalização Intercelular , Lectinas/biossíntese , Lectinas/genética , Camundongos , Fator de Crescimento Neural/biossíntese , Fator de Crescimento Neural/genética , Proteínas/genética , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , RNA Viral/biossíntese , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Crescimento Transformador beta/metabolismo , Infecções Tumorais por Vírus/patologia , Infecções Tumorais por Vírus/virologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas do Envelope Viral/biossíntese , Proteínas do Envelope Viral/genética , beta-N-Acetil-Hexosaminidases/biossíntese , beta-N-Acetil-Hexosaminidases/genética , Receptor de Interferon gamaRESUMO
The etiology of idiopathic pulmonary fibrosis (IPF) is unknown. Because viral pathogenesis of IPF has been suggested, we have established a murine model of progressive pulmonary fibrosis by infecting IFN-gammaR-deficient mice (IFN-gammaR(-/-)) with the murine gamma-herpesvirus 68. Because alveolar macrophages in humans with IPF have been implicated in driving the profibrotic response, we studied their role in our model. Chronic herpesvirus infection of the lung was associated with recruitment of alveolar macrophages to areas with epithelial hyperplasia and fibrosis in infected lungs. Using immunohistochemistry, Western blot, and RT-PCR techniques, we demonstrated that recruited alveolar macrophages showed high levels of expression of the proteins Ym1/2, FIZZ1 (found in inflammatory zone 1), insulin-like growth factor-1, and arginase I, and also active transcription of fibronectin, indicative of activation of macrophages by an alternative pathway. Arginase I expression was also evident in interstitial fibroblasts, and increased arginase activity was found in lungs of infected animals. Lung tissue from patients with IPF showed increased expression of arginase I in epithelial cells, fibroblast foci, and alveolar macrophages compared with normal lung. These results suggest that virus-induced upregulation of arginase I could be a mechanism driving lung fibrogenesis.
Assuntos
Gammaherpesvirinae , Infecções por Herpesviridae/imunologia , Pulmão/enzimologia , Ativação de Macrófagos/imunologia , Macrófagos Alveolares/fisiologia , Fibrose Pulmonar/imunologia , Receptores de Interferon/genética , Animais , Arginase/metabolismo , Doença Crônica , Fibronectinas/metabolismo , Infecções por Herpesviridae/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células NIH 3T3 , Fibrose Pulmonar/enzimologia , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/virologia , Transdução de Sinais , Transfecção , Receptor de Interferon gamaRESUMO
The NF-kappaB/Rel transcription factor family plays a central role in coordinating the expression of a variety of genes that regulate stress responses, immune cell activation, apoptosis, proliferation, differentiation, and oncogenic transformation. Interventions that target the NF-kappaB pathway may be therapeutic for a variety of pathologies, especially immune/inflammatory diseases. Using membrane translocating sequence (MTS) technology, we developed a cell-permeable dominant inhibitor of NF-kappaB activation, termed IkappaBalpha-(DeltaN)-MTS. This molecule contains a 12-amino acid MTS motif attached to the COOH-terminal region of a nondegradable inhibitor protein [IkappaBalpha-(DeltaN)]. The recombinant protein enters cells and localizes in the cytoplasm. Delivery of the IkappaBalpha-(DeltaN)-MTS to cell lines and primary cells inhibited nuclear translocation of NF-kappaB proteins induced by cell activation. The protein also effectively inhibited NF-kappaB activation in vivo in two different animal models: NF-kappaB activation in response to skin wounding in mice and NF-kappaB activation in lungs after endotoxin treatment in sheep. Inhibition of NF-kappaB by the IkappaBalpha-(DeltaN)-MTS in the endotoxin model attenuated physiological responses to endotoxemia. These data demonstrate that activation of NF-kappaB can be inhibited using a recombinant protein designed to penetrate into cells. This technology may provide a new approach to NF-kappaB pathway-targeted therapies.
Assuntos
Proteínas I-kappa B/farmacocinética , Pneumopatias/tratamento farmacológico , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Proteínas Recombinantes de Fusão/farmacocinética , Animais , Divisão Celular/imunologia , Permeabilidade da Membrana Celular , Sistemas de Liberação de Medicamentos , Endotoxemia/tratamento farmacológico , Endotoxemia/imunologia , Expressão Gênica , Humanos , Proteínas I-kappa B/genética , Pneumopatias/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Inibidor de NF-kappaB alfa , Células NIH 3T3 , Proteínas Recombinantes de Fusão/genética , Ovinos , Linfócitos T/efeitos dos fármacos , Timo/citologiaRESUMO
We sought to determine whether an intact bone marrow is essential to lung repair following bleomycin-induced lung injury in mice, and the mechanisms of any protective effects conferred by bone marrow-derived mesenchymal stem cell (BMDMSC) transfer. We found that myelosupression increased susceptibility to bleomycin injury and that BMDMSC transfer was protective. Protection was associated with the differentiation of engrafted BMDMSC into specific and distinct lung cell phenotypes, with an increase in circulating levels of G-CSF and GM-CSF (known for their ability to promote the mobilization of endogenous stem cells) and with a decrease in inflammatory cytokines. In vitro, cells from injured, but not from normal, mouse lung produced soluble factors that caused BMDMSC to proliferate and migrate toward the injured lung. We conclude that bone marrow stem cells are important in the repair of bleomycin-injured lung and that transfer of mesenchymal stem cells protects against the injury. BMDMSC localize to the injured lung and assume lung cell phenotypes, but protection from injury and fibrosis also involves suppression of inflammation and triggering production of reparative growth factors.